title: Instant super-resolution imaging in live cells and embryos via analog image processing.
abstract: Existing super-resolution fluorescence microscopes compromise acquisition speed to provide subdiffractive sample information. We report an analog implementation of structured illumination microscopy that enables three-dimensional (3D) super-resolution imaging with a lateral resolution of 145 nm and an axial resolution of 350 nm at acquisition speeds up to 100 Hz. By using optical instead of digital image-processing operations, we removed the need to capture, store and combine multiple camera exposures, increasing data acquisition rates 10- to 100-fold over other super-resolution microscopes and acquiring and displaying super-resolution images in real time. Low excitation intensities allow imaging over hundreds of 2D sections, and combined physical and computational sectioning allow similar depth penetration to spinning-disk confocal microscopy. We demonstrate the capability of our system by imaging fine, rapidly moving structures including motor-driven organelles in human lung fibroblasts and the cytoskeleton of flowing blood cells within developing zebrafish embryos.
authors.affiliation: Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Andrew G
authors.initials: AG
authors.lastname: York
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Chandris
authors.forename: Damian Dalle
authors.initials: DD
authors.lastname: Nogare
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Head
authors.forename: Peter
authors.initials: P
authors.lastname: Wawrzusin
authors.forename: Robert S
authors.initials: RS
authors.lastname: Fischer
authors.forename: Ajay
authors.initials: A
authors.lastname: Chitnis
authors.forename: Hari
authors.initials: H
authors.lastname: Shroff
mesh_terms: D000818:Animals; D004622:Embryo, Mammalian / Q000166:cytology*; D008856:Microscopy, Fluorescence